Fig. 3. Differential P2X7R-dependent responses in GBM primary cell lines.

a, b Representative traces showing BzATP -dependent cytosolic Ca²⁺ increase in the absence or presence of 5 min pretreatment with AZ10606120 (1 μM). c, d Representative traces showing ethidium bromide uptake following stimulation with increasing concentrations of BzATP alone or applied after 5 min pretreatment with AZ10606120 (1 μM). e, f BzATP-dependent cytotoxicity in GBM cell lines in the absence or presence of P2X7R antagonists AZ10606120 or A740003, measured as percent LDH release normalized to the total amount of protein (µg). Data are shown as mean ± SEM; n = 3. Statistical analyses were conducted with unpaired two-tailed Student’s t-test. g mRNA expression levels of P2X7A and P2X7B isoforms in GB40 and GB48 cells. Data were normalized to HPRT-1 and GAPDH housekeeping genes. Data are shown as mean ± SEM; n = 3. Statistical analyses were conducted with unpaired two-tailed Student’s t-test. h Percentage of surviving cells measured by soft agar assay at 28 days from radiation treatment at different doses in GB40 and GB48. Data are shown as the mean ± SEM; n = 6. Statistical analyses were conducted with unpaired two-tailed Student’s t-test. P values: n.s not significant; *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001.